Sundin Bryan A, Chiu Chun-Hwei, Riffle Michael, Davis Trisha N, Muller Eric G D
Department of Biochemistry, University of Washington, Seattle, WA 98195, USA.
Yeast. 2004 Jul 15;21(9):793-800. doi: 10.1002/yea.1133.
The localization of proteins can give important clues about their function and help sort data from large-scale proteomic screens. Forty-five proteins were tagged with the GFP variant YFP. These proteins were chosen because they are encoded by genes that display strong cell cycle-dependent expression that peaks in G(1). Most of these proteins localize to either the nucleus or to sites of cell growth. We are able to assign new cellular component GO terms to ASF2, TOS4, RTT109, YBR070C, YKR090W, YOL007C, YOL019W and YPR174C. We also have localization data for 21 other proteins. Noteworthy localizations were found for Rfa1p, a member of the DNA replication A complex, and Pri2p and Pol12p, subunits of the alpha-DNA polymerase : primase complex. In addition to its nuclear localization, Rfa1p assembled into cytoplasmic foci adjacent to the nucleus in cells during the G(1)-S phase transition of the cell cycle. Pri2 and Pol12 took on a beaded appearance at the G(1)-S transition and later in the cell cycle were enriched in the nuclear envelope. A new spindle pole body/nuclear envelope component encoded by YPR174 was identified. The cell cycle-dependent abundance of Tos4p mirrored Yox1p and these two proteins were the only proteins that were found exclusively at the G(1)-S phase of the cell cycle. A complete list of localizations, along with images, can be found at our website (http://www.yeastrc.org/cln2/).
蛋白质的定位能够为其功能提供重要线索，并有助于整理大规模蛋白质组学筛选的数据。45种蛋白质用绿色荧光蛋白变体黄色荧光蛋白（GFP variant YFP）进行了标记。选择这些蛋白质是因为它们由在G(1)期达到峰值的具有强烈细胞周期依赖性表达的基因编码。这些蛋白质中的大多数定位于细胞核或细胞生长部位。我们能够为ASF2、TOS4、RTT109、YBR070C、YKR090W、YOL007C、YOL019W和YPR174C赋予新的细胞成分基因本体术语（GO terms）。我们还有另外21种蛋白质的定位数据。发现DNA复制A复合物成员Rfa1p以及α-DNA聚合酶：引发酶复合物的亚基Pri2p和Pol12p有值得注意的定位。除了其核定位外，在细胞周期的G(1)-S期转变过程中，Rfa1p在细胞核附近的细胞质中聚集成灶。Pri2和Pol12在G(1)-S期转变时呈现串珠状外观，并且在细胞周期后期在核膜中富集。鉴定出了由YPR174编码的一种新的纺锤体极体/核膜成分。Tos4p的细胞周期依赖性丰度反映了Yox1p，并且这两种蛋白质是仅在细胞周期的G(1)-S期发现的唯一蛋白质。定位的完整列表以及图像可在我们的网站（http://www.yeastrc.org/cln2/）上找到。
